Compositions and methods for treatment of gaucher disease

Abstract

Described herein are methods for treating a subject having or at risk of developing Gaucher disease, by administering one or more agents that increase expression and/or activity of glucocerebrosidase (GBA) and/or scavenger receptor class B member 2 (SCARB2), such as pluripotent cells that express GBA and/or SCARB2, and to the subject. Also disclosed are compositions comprising one or more agents that increase the expression and/or activity of GBA and/or SCARB2.

1 . A method of treating a subject diagnosed as having or at risk of developing Gaucher disease, the method comprising providing to the subject one or more agents that collectively increase expression and/or activity of β-glucocerebrosidase (GBA) and scavenger receptor class B member 2 (SCARB2). 
     
     
         2 . The method of  claim 1 , wherein the one or more agents comprise a first agent that increases expression and/or activity of GBA and a second agent that increases expression and/or activity of SCARB2; optionally, wherein the first agent comprises (i) one or more polynucleotides comprising a transgene that encodes a GBA protein, (ii) one or more interfering RNA (RNAi) molecules that collectively increase expression and/or activity of the GBA protein, (iii) one or more polynucleotides encoding the one or more RNAi molecules that collectively increase expression and/or activity of the GBA protein, (iv) a GBA protein, or (v) one or more small molecules that collectively increase expression and/or activity of the GBA protein, and wherein the second agent comprises (vi) one or more polynucleotides comprising a transgene that encodes a SCARB2 protein, (vii) one or more RNAi molecules that collectively increase expression and/or activity of the SCARB2 protein, (viii) one or more polynucleotides encoding the one or more RNAi molecules that collectively increase expression and/or activity of the SCARB2 protein, (ix) a SCARB2 protein, (x) one or more small molecules that collectively increase expression and/or activity of the SCARB2 protein. 
     
     
         3 . The method of  claim 2 , wherein the GBA protein has an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO: 1. 
     
     
         4 . The method of  claim 3 , wherein the GBA protein has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 1; optionally, wherein the GBA has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 1; optionally, wherein the GBA has the amino acid sequence of SEQ ID NO: 1. 
     
     
         5 . The method of  claim 2 , wherein the GBA has an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO: 5. 
     
     
         6 . The method of  claim 5 , wherein the GBA has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 5; optionally, wherein the GBA has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 5; optionally; wherein the GBA has the amino acid sequence of SEQ ID NO: 5. 
     
     
         7 . The method of any one of  claims 2-6  wherein the transgene encoding GBA has a nucleic acid sequence that is at least 85% identical to the nucleic acid sequence of SEQ ID NO: 6. 
     
     
         8 . The method of  claim 7 , wherein the transgene encoding GBA has a nucleic acid sequence that is at least 90% identical to the nucleic acid sequence of SEQ ID NO: 6; optionally, wherein the transgene encoding GBA has a nucleic acid sequence that is at least 95% identical to the nucleic acid sequence of SEQ ID NO: 6; optionally, wherein the transgene encoding GBA has the nucleic acid sequence of SEQ ID NO: 6. 
     
     
         9 . The method of any one of  claims 2-6 , wherein the transgene encoding GBA has a nucleic acid sequence that is at least 85% identical to the nucleic acid sequence of SEQ ID NO: 7. 
     
     
         10 . The method of  claim 9 , wherein the transgene encoding GBA has a nucleic acid sequence that is at least 90% identical to the nucleic acid sequence of SEQ ID NO: 7; optionally, wherein the transgene encoding GBA has a nucleic acid sequence that is at least 95% identical to the nucleic acid sequence of SEQ ID NO: 7; optionally, wherein the transgene encoding GBA has the nucleic acid sequence of SEQ ID NO: 7. 
     
     
         11 . The method of any one of  claims 2-6 , wherein the transgene encoding GBA has a nucleic acid sequence that is at least 85% identical to the nucleic acid sequence of SEQ ID NO: 11. 
     
     
         12 . The method of  claim 11 , wherein the transgene encoding GBA has a nucleic acid sequence that is at least 90% identical to the nucleic acid sequence of SEQ ID NO: 11; optionally, wherein the transgene encoding GBA has a nucleic acid sequence that is at least 95% identical to the nucleic acid sequence of SEQ ID NO: 11; optionally, wherein the transgene encoding GBA has the nucleic acid sequence of SEQ ID NO: 11. 
     
     
         13 . The method of any one of  claims 2-12 , wherein the GBA comprises a signal peptide, wherein the signal peptide has an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO: 12. 
     
     
         14 . The method of  claim 13 , wherein the signal peptide has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 12; optionally, wherein the signal peptide has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 12; optionally, wherein the signal peptide has the amino acid sequence of SEQ ID NO: 12. 
     
     
         15 . The method of  claim 2-12 , wherein the GBA comprises a signal peptide, wherein the signal peptide is encoded by a polynucleotide that is at least 85% identical to the nucleic acid sequence of SEQ ID NO: 13. 
     
     
         16 . The method of  claim 15 , wherein the signal peptide is encoded by a polynucleotide that is at least 90% identical to the nucleic acid sequence of SEQ ID NO: 13; optionally, wherein the signal peptide is encoded by a polynucleotide that is at least 95% identical to the nucleic acid sequence of SEQ ID NO: 13; optionally, wherein the signal peptide is encoded by a polynucleotide having the nucleic acid sequence of SEQ ID NO: 13. 
     
     
         17 . The method of any one of  claims 2-16 , wherein the SCARB2 has an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO: 14. 
     
     
         18 . The method of  claim 17 , wherein the SCARB2 has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 14; optionally, wherein the SCARB2 has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 14; optionally, wherein the SCARB2 has an amino acid sequence of SEQ ID NO: 14. 
     
     
         19 . The method of claim any one of  claims 2-16 , wherein the SCARB2 has an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO: 15. 
     
     
         20 . The method of  claim 19 , wherein the SCARB2 has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 15; optionally, wherein the SCARB2 has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 15; optionally, wherein the SCARB2 has an amino acid sequence of SEQ ID NO: 15. 
     
     
         21 . The method of claim of any one of  claims 2-20 , wherein the polynucleotide encoding SCARB2 has a nucleic acid sequence that is at least 85% identical to the nucleic acid sequence of SEQ ID NO: 16. 
     
     
         22 . The method of  claim 21 , wherein the polynucleotide encoding SCARB2 has a nucleic acid sequence that is at least 90% identical to the nucleic acid sequence of SEQ ID NO: 16; optionally, wherein the polynucleotide encoding SCARB2 has a nucleic acid sequence that is at least 95% identical to the nucleic acid sequence of SEQ ID NO: 16; optionally, wherein the polynucleotide encoding SCARB2 has a nucleic acid sequence of SEQ ID NO: 16. 
     
     
         23 . The method of any one of  claims 2-20 , wherein the polynucleotide encoding SCARB2 has a nucleic acid sequence that is at least 85% identical to the nucleic acid sequence of SEQ ID NO: 17. 
     
     
         24 . The method of  claim 23 , wherein the polynucleotide encoding SCARB2 has a nucleic acid sequence that is at least 90% identical to the nucleic acid sequence of SEQ ID NO: 17; optionally, wherein the polynucleotide encoding SCARB2 has a nucleic acid sequence that is at least 95% identical to the nucleic acid sequence of SEQ ID NO: 17; optionally, wherein the polynucleotide encoding SCARB2 has a nucleic acid sequence of SEQ ID NO: 17. 
     
     
         25 . The method of  claim 2-24 , wherein the SCARB2 comprises a signal peptide, wherein the signal peptide has an amino acid sequence that is at least 70% identical to the amino acid sequence of SEQ ID NO: 49; optionally, wherein the signal peptide has an amino acid sequence that is at least 75% identical to the amino acid sequence of SEQ ID NO: 49; optionally, wherein the signal peptide has an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 49; optionally, wherein the signal peptide has an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO: 49; optionally, wherein the signal peptide has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 49; optionally, wherein the signal peptide has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 49; optionally, wherein the signal peptide has an amino acid sequence of SEQ ID NO: 49. 
     
     
         26 . The method of any one of  claims 2-25 , wherein the SCARB2 is a GBA-binding domain of SCARB2. 
     
     
         27 . The method of  claim 26 , wherein the GBA-binding domain of the SCARB2 protein has an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO: 18; optionally, wherein the GBA-binding domain of the SCARB2 protein has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 18; optionally, wherein the GBA-binding domain of the SCARB2 protein has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 18; optionally, wherein the GBA-binding domain of the SCARB2 protein has an amino acid sequence of SEQ ID NO: 18. 
     
     
         28 . The method of  claim 26 , wherein the GBA-binding domain of the SCARB2 protein has an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO: 19; optionally, wherein the GBA-binding domain of the SCARB2 protein has an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 19; optionally, wherein the GBA-binding domain of the SCARB2 protein has an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 19; optionally, wherein the GBA-binding domain of the SCARB2 protein has an amino acid sequence of SEQ ID NO: 19. 
     
     
         29 . The method of claim any one of  claims 2-28  wherein the GBA and/or SCARB2 is a fusion protein comprising GBA or SCARB2 and a glycosylation independent lysosomal targeting (GILT) tag. 
     
     
         30 . The method of  claim 29 , wherein the GILT tag comprises a human IGF-II mutein having an amino acid sequence that is at least 70% identical to the amino acid sequence of mature human IGF-II (SEQ ID NO: 22), and having diminished binding affinity for the insulin receptor relative to the affinity of naturally-occurring human IGF-II for the insulin receptor, wherein the IGF-II mutein is resistant to furin cleavage and binds to the human cation-independent mannose-6-phosphate receptor in a mannose-6-phosphate-independent manner. 
     
     
         31 . The method of  claim 30 , wherein the IGF-II mutein comprises a mutation within a region corresponding to amino acids 30-40 of SEQ ID NO: 22, and wherein the mutation abolishes at least one furin protease cleavage site. 
     
     
         32 . The method of  claim 31 , wherein the mutation is an amino acid substitution, deletion, and/or insertion. 
     
     
         33 . The method of  claim 32 , wherein the mutation is an Ala amino acid substitution at a position corresponding to Arg37 of SEQ ID NO: 22. 
     
     
         34 . The method of  claim 32 , wherein the mutation is a deletion or replacement of amino acid residues corresponding to positions selected form the group consisting of 31-40, 32-40, 33-40, 34-40, 30-39, 31-39, 32-39, 34-37, 33-39, 35-39, 36-39, 37-40, 34-40 of SEQ ID NO: 22, and combinations thereof. 
     
     
         35 . The method of any one of  claims 29-34 , wherein the GBA fusion protein and/or the SCARB2 fusion protein comprises a receptor-binding (Rb) domain of apolipoprotein E (ApoE). 
     
     
         36 . The method of  claim 35 , wherein the Rb domain comprises a portion of ApoE having the amino acid sequence of residues 25-185, 50-180, 75-175, 100-170, 125-160, or 130-150 of SEQ ID NO: 29. 
     
     
         37 . The method of  claim 35 , wherein the Rb domain comprises a region having at least 70% sequence identity to the amino acid sequence of residues 159-167 of SEQ ID NO: 29. 
     
     
         38 . The method of claim any one of  claims 2-37  wherein the GBA protein or the SCARB2 protein is a fusion protein comprising GBA or SCARB2 and a cell-penetrating peptide (CPP). 
     
     
         39 . The method of  claim 38 , wherein the CPP has an amino acid sequence having at least 85% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 30-48; optionally, wherein the CPP has an amino acid sequence having at least 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 30-48; optionally, wherein the CPP has an amino acid sequence having at least 95% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 30-48; optionally, wherein the CPP has an amino acid sequence having the amino acid sequence of any one of SEQ ID NOs: 30-48. 
     
     
         40 . The method of any one of  claims 2-39 , wherein the transgene encoding GBA and/or SCARB2 further comprises a microRNA (miRNA) targeting sequence in the 3′-UTR. 
     
     
         41 . The method of  claim 40 , wherein the miRNA targeting sequence is a miR-126 targeting sequence. 
     
     
         42 . The method of any one of  claims 2-41 , wherein the GBA and/or the SCARB2 penetrates the blood brain barrier (BBB) in the subject. 
     
     
         43 . The method of any one of  claims 2-42 , wherein the one or more RNAi molecules comprise short interfering RNA (siRNA), short hairpin RNA (shRNA), and/or microRNA (miRNA). 
     
     
         44 . The method of any one of  claims 2-43 , wherein the one or more polynucleotides are provided to the subject by administering to the subject a composition comprising a population of cells that together contain nucleic acids encoding the GBA and/or SCARB2 protein. 
     
     
         45 . The method of  claim 44 , wherein the population is a uniform population of cells that contain nucleic acids encoding the proteins or a heterogeneous population of cells that together contain nucleic acids encoding the GBA and/or SCARB2 protein. 
     
     
         46 . The method of  claim 44 or 45 , wherein the cells are pluripotent cells or multipotent cells. 
     
     
         47 . The method of  claim 46 , wherein the multipotent cells are CD34+ cells. 
     
     
         48 . The method of  claim 47 , wherein the CD34+ cells are hematopoietic stem cells (HSCs) or myeloid progenitor cells (MPCs). 
     
     
         49 . The method of  claim 46 , wherein the pluripotent cells are embryonic stem cells (ESCs) or induced pluripotent stem cells (iPSCs). 
     
     
         50 . The method of  claim 44 or 45 , wherein the cells are blood lineage progenitor cells (BLPCs), microglial progenitor cells, monocytes, macrophages, or microglia. 
     
     
         51 . The method of  claim 50 , wherein the BLPCs are monocytes. 
     
     
         52 . The method of any one of  claims 1-51 , wherein a population of endogenous hematopoietic cells in the subject has been ablated prior to administration of the one or more agents to the subject, optionally wherein the hematopoietic cells are CD34+ cells, BLPCs, microglial progenitor cells, monocytes, macrophages, or microglia. 
     
     
         53 . The method of any one of  claims 1-51 , the method comprising ablating a population of endogenous hematopoietic cells in the subject prior to administering the one or more agents to the subject, optionally wherein the hematopoietic cells are CD34+ cells, BLPCs, microglial progenitor cells, monocytes, macrophages, or microglia. 
     
     
         54 . The method of  claim 52 or 53 , wherein the microglia are ablated using an agent selected from the group consisting of busulfan, PLX3397, PLX647, PLX5622, treosulfan, and clodronate liposomes, by radiation therapy, or a combination thereof. 
     
     
         55 . The method of any one of  claims 1-54 , wherein the one or more agents is administered systemically to the subject. 
     
     
         56 . The method of  claim 55 , wherein the one or more agents is administered to the subject by way of intravenous injection. 
     
     
         57 . The method of any one of  claims 1-54 , wherein the one or more agents is administered directly to the central nervous system of the subject. 
     
     
         58 . The method of  claim 57 , wherein the one or more agents is administered to the subject by way of intracerebroventricular injection, stereotactic injection, or a combination thereof. 
     
     
         59 . The method of any one of  claims 1-54 , wherein the one or more agents is administered directly to the bone marrow of the subject. 
     
     
         60 . The method of  claim 59 , wherein the one or more agents is administered to the subject by way of intraosseous injection. 
     
     
         61 . The method of any one of  claims 44-60 , wherein the cells are autologous cells or allogeneic cells. 
     
     
         62 . The method of any one of  claims 44-61 , wherein the cells are transfected or transduced ex vivo to express the GBA and/or SCARB2. 
     
     
         63 . The method of  claim 62 , wherein the cells are transduced with a viral vector selected from the group consisting of an adeno-associated virus (AAV), an adenovirus, a parvovirus, a coronavirus, a rhabdovirus, a paramyxovirus, a picornavirus, an alphavirus, a herpes virus, a poxvirus, and a Retroviridae family virus. 
     
     
         64 . The method of  claim 63 , wherein the viral vector is a Retroviridae family viral vector. 
     
     
         65 . The method of  claim 64 , wherein the Retroviridae family viral vector is a lentiviral vector. 
     
     
         66 . The method of  claim 64 , wherein the Retroviridae family viral vector is an alpharetroviral vector. 
     
     
         67 . The method of  claim 64 , wherein the Retroviridae family viral vector is a gammaretroviral vector. 
     
     
         68 . The method of any one of  claims 63-67 , wherein the Retroviridae family viral vector comprises a central polypurine tract, a woodchuck hepatitis virus post-transcriptional regulatory element, a 5′-LTR, HIV signal sequence, HIV Psi signal 5′-splice site, delta-GAG element, 3′-splice site, and a 3′-self inactivating LTR. 
     
     
         69 . The method of  claim 63 , wherein the viral vector is a pseudotyped viral vector. 
     
     
         70 . The method of  claim 69 , wherein the pseudotyped viral vector selected from the group consisting of a pseudotyped AAV, a pseudotyped adenovirus, a pseudotyped parvovirus, a pseudotyped coronavirus, a pseudotyped rhabdovirus, a pseudotyped paramyxovirus, a pseudotyped picornavirus, a pseudotyped alphavirus, a pseudotyped herpes virus, a pseudotyped poxvirus, and a pseudotyped Retroviridae family virus. 
     
     
         71 . The method of any one of  claims 62-70 , wherein the pluripotent cells are transduced to express the GBA and SCARB2 from separate monocistronic expression cassettes. 
     
     
         72 . The method of any one of  claims 62-70 , wherein the pluripotent cells are transduced to express the GBA and SCARB2 from a polycistronic expression cassette. 
     
     
         73 . The method of  claim 72 , wherein the pluripotent cells are transduced to express the GBA and SCARB2 from a bicistronic expression cassette. 
     
     
         74 . The method of  claim 72 or 73 , wherein the polycistronic expression cassette comprises an internal ribosomal entry site (IRES) positioned between a polynucleotide encoding the GBA and a polynucleotide encoding the SCARB2. 
     
     
         75 . The method of  claim 72 or 73 , wherein the polycistronic expression cassette comprises a 2A polynucleotide positioned between a polynucleotide encoding the GBA and a polynucleotide encoding the SCARB2. 
     
     
         76 . The method of  claim 75 , wherein the 2A polynucleotide comprises a F2A, P2A, E2A, or T2A polynucleotide. 
     
     
         77 . The method of any one of  claims 2-76 , wherein one or more of the polynucleotides comprises a transgene encoding one or more of the proteins operably linked to a ubiquitous promoter, a cell lineage-specific promoter, or a synthetic promoter. 
     
     
         78 . The method of  claim 77 , wherein the ubiquitous promoter is selected from the group consisting of an elongation factor 1-alpha (EF1α) promoter, phosphoglycerate kinase 1 (PGK) promoter, or EF1α promoter containing elements of locus control region of the β-globin gene containing regions of erythroid-specific DNase I hypersensitivity (HS) regions 2, 3, and 4 (β-LCR(HS4,3,2)-EFS promoter). 
     
     
         79 . The method of  claim 77 , wherein the cell lineage-specific promoter is selected from the group consisting of a CD68 molecule (CD68) promoter, CD11 b molecule (CD11b) promoter, C-X3-C motif chemokine receptor 1 (CX3CR1) promoter, allograft inflammatory factor 1 promoter (AIF1) promoter, purinergic receptor P2Y12 (P2Y12) promoter, transmembrane protein 119 (TMEM119) promoter, or colony stimulating factor 1 receptor (CSF1R) promoter. 
     
     
         80 . The method of  claim 77 , wherein the synthetic promoter is a Myeloproliferative Sarcoma Virus Enhancer, Negative Control Region Deleted, dl587rev Primer-Binding Site Substituted (MND) promoter. 
     
     
         81 . The method of  claim 80 , wherein the MND promoter comprises a polynucleotide having at least 85% sequence identity to the nucleic acid sequence of SEQ ID NO: 20. 
     
     
         82 . The method of  claim 80 , wherein the MND promoter comprises a polynucleotide having at least 85% sequence identity to the nucleic acid sequence of SEQ ID NO: 21. 
     
     
         83 . The method of any one of  claims 1-82 , wherein the Gaucher disease is associated with one or more mutations in the GBA gene. 
     
     
         84 . The method of  claim 83 , wherein the one or more mutations in the GBA gene comprise a p.N370S substitution, p.R463C substitution, p.L444P substitution, p.D409H substitution, p.R463C substitution, p.R496H substitution, p. F252I substitution, p.A456P substitution, p.V460V substitution, p.V394L, p.E326K substitution, p.G377S substitution, p.N188S substitution, c.84insG insertion, c.84dupG (84GG) duplication, c.115+1 G>A substitution, or c.IVS2DS+1 G-A splice site mutation. 
     
     
         85 . The method of any one of  claims 1-84 , wherein the Gaucher disease is Type 1 Gaucher disease. 
     
     
         86 . The method of  claim 85 , wherein the subject has a confirmed diagnosis of Type 1 Gaucher disease based on genotyping, deficient GBA activity in the blood of the subject, and/or clinical phenotype. 
     
     
         87 . The method of  claim 86 , wherein the deficient GBA activity in the subject is defined as activity that is equal to or greater than 15% of activity of GBA in a control reference patient not diagnosed as having Gaucher disease. 
     
     
         88 . The method of any one of  claims 1-84 , wherein the Gaucher disease is Type 2 Gaucher disease. 
     
     
         89 . The method of any one of  claims 1-84 , wherein the Gaucher disease is Type 3 Gaucher disease. 
     
     
         90 . The method of any one of  claims 1-89 , wherein the Gaucher disease is associated with one or more mutations in the SCARB2 gene. 
     
     
         91 . The method of  claim 90 , wherein the one or more mutations in the SCARB2 gene comprise a p.Q471G substitution, p.H363N substitution, p.Q288Ter nonsense mutation, p.W178Ter nonsense mutation, p.W146fs frameshift mutation, p.Glu420fs frameshift mutation, g.76168478T>G transversion mutation, g.1239+1G-T splice site mutation, or g.76168401dup splice site mutation p.Q288Ter nonsense mutation, p.W178Ter nonsense mutation, p.W146fs frameshift mutation, p.Glu420fs frameshift mutation, g.76168478T>G transversion mutation, g.1239+1G-T splice site mutation, or g.76168401dup splice site mutation. 
     
     
         92 . The method of any one of  claims 1-91 , wherein the subject is a human. 
     
     
         93 . The method of any one of  claims 1-92 , wherein the subject has undergone enzyme replacement therapy (ERT) comprising a total monthly dose of GBA ERT that is greater than 30 U/kg and less than 120 U/kg for 24 or more consecutive months at a time of treatment with the one or more agents. 
     
     
         94 . The method of  claim 93 , wherein the subject has received a biweekly dose of GBA ERT greater than or equal to 15 U/kg and less than or equal to 60 U/kg. 
     
     
         95 . The method of  claim 93 , wherein the subject has received a weekly dose of GBA ERT greater than or equal to 7.5 U/kg and less than or equal to 30 U/kg. 
     
     
         96 . The method of any one of  claims 1-95 , wherein the subject has received substrate reduction therapy (SRT) for Gaucher disease during the 24 months immediately preceding treatment with the one or more agents. 
     
     
         97 . The method of any one of  claims 1-95 , wherein the subject has not received SRT for Gaucher disease during the 24 months immediately preceding treatment with the one or more agents.